Following the death of Robert Wenthold at the end of October, the lab began a plan to coordinate the systematic break-up of the lab: 1) facilitating the movement of many of the researchers to other labs or positions over the next few months;2) helping other researchers to continue and finish projects in the lab;and 3) overseeing the moving and clean-up of the lab benches, instruments, refrigerators and freezers, and desk areas. Most of this was completed by the end of April, 2010. After that, the lab continued to support the projects of some of the researchers who had moved to the lab of Dr. Katherine Roche in NINDS, and more extensively, the projects of those who continued to maintain closer ties with the lab, including Drs. Martin Horak, Chan-Ying Zheng, Philip Wang, and Gail Seabold. The lab continued doing weekly neuron cell culture preparations through the early summer and now does them sporadically as need by these researchers. Basically, work now is done as collaborations with PIs of other labs. Much of this continues the work done in the lab of Dr. Wenthold on the trafficking of glutamate receptors and associated proteins of the neuronal synapse, including some of the projects mentioned in his 2009 Annual Report. One of the major concentrations of this current project is to determine the structural and functional arrangement of scaffolding and cytoskeletal proteins associated with glutamate receptors in the postsynaptic density of synapses;this is crucial to understanding how synapses work and change to mediate learning and memory. In addition to these general central synaptic studies, the lab collaborates on studies in the auditory system. Recently the lab has become involved in studies of development and pathology of the cochlea, and in the near future also hopes to develop some work on central auditory function (which was a one of the major specialties of the lab in earlier years). The specific research projects that the lab has been involved in as collaborators this year are as follows: With researchers of the original Wenthold lab: 1) Completion and publication of a paper on LM and EM studies on extrasynaptic NMDA receptors, published in the journal Neuroscience. 2) Completion and publication of a paper with Chan-Ying Zheng on PSD-95 and SAP102 in synaptic spines, published in the Journal of Neuroscience. 3) Completion of a study with Philip Wang and Stephan Brenowitz on NMDA receptors on neuronal growth cones, recently submitted. Performed EM analyses used in this study and also provided the cultured neurons. 4) Continued work on a SALM1 paper with Gail Seabold and Philip Wang. This is an adhesion protein associated with glutamate receptors at synapses. This paper is nearly complete. Performed EM analyses used in this study and provided the cultured neurons. Also supervised a summer student this summer, Arthur Zhou (hired by Dr. Kachar), who performed LM confocal microscopy as part of this study. 5) Continued work on EM immunogold studies as part of a larger study in progress with Chan-Ying Zheng and Bechara Kachar (i.e., the structural study of scaffolding and cytoskeleton associated with the postsynaptic density of synapses, as noted above). Also supervised a summer student this summer, Arthur Zhou (hired by Dr. Kachar), who performed analyses on the EM data of the lab, as part of this study. 6) Working with Chan-Ying Zheng to complete a video method paper on FRAP of synaptic spines for the journal JoVE. 7) Working with Chan-Ying Zheng to complete a paper as an addendum to the published paper on PSD-95 and SAP102, to be published in the journal Communicative &Integrative Biology. 8) The lead investigator was invited to write a review paper on MAGUK plasticity for The Neuroscientist, and enlisted the help of Chan-Ying Zheng, Gail Seabold and Martin Horak. This just was accepted for publication. With others at NIDCD: 1) Worked with Drs. Dabin Ren and Xin-Xing Gu to complete 2 separate EM studies of immunogold localization on bacteria associated with Otitis Media. 2) Working with researchers from the lab of Dr. Andy Griffith on ultrastructural changes in the cochlea associated with mutants associated with deafness. This includes cutting sections and doing EM for this and also training Kyu-Yup Lee to do EM for this, and helping with the analyses and continuing EM studies. 3) Training and working with Kate Szarama from the labs of Drs. Matt Kelley and Richard Chadwick looking at the ultrastructural changes in the cochlea related to disease. 4) Working, as already noted above, on 4 projects/papers with Chan-Ying Zheng as now part of the projects of the lab of Dr. Kachar. 5) Involved in some preliminary studies of EM immunogold distribution with researchers (esp. Jonathan Bird) in the lab of Dr. Tom Friedman. Also, trained Inna Bellyantseva on EM technique. With other NIH institutes: 1) Completed a study on postsynaptic SNAP23 with Dr. Katherine Roche of NINDS and Dr. Paul Roche of NCI;this was published in Nature Neuroscience in the spring. 2) Working on several studies of EM localization of sonic hedgehog and its receptors, patched and smoothened, with Drs. Pamela Yao and Mark Mattson of NIA. The first paper is in the process of submission. 3) Involved in 2 preliminary studies on mutations of proteins that affect the nervous system (including a mouse model for schizophrenia) with Drs. Jie-Min Jia and Song Jiao of NIMH. 4) Worked extensively in the spring with Dr. Brian Mozer of NHLBI in a study of an adhesion protein in Drosophila synapses. With labs outside of NIH: 1) Working with Dr. Jinhyun Kim of Janelia Farm Research Campus (HHMI), on EM immunolocalization of a protein in the synaptic cleft. 2) Working on EM immunogold contributions to 3 papers related to pain effects on neuronal plasticity of the spinal cord, currently in preparation or submission to journals with Dr. Yuan-Xiang Tao of Johns Hopkins School of Medicine in Baltimore;these involve other collaborators such as Drs. Richard Huganir and Jeffrey Rothstein of Johns Hopkins, and D. Olga Kopach of the Bogomoletz Institute of Physiology in the Ukraine. 3) Working on an EM immungold study of the brain of AMPA receptor knock-in mice with researchers in the laboratory of Richard Huganir of Johns Hopkins School of Medicine in Baltimore. 4) Working with researchers in the laboratory of Dr. Paul Worley of Johns Hopkins School of Medicine in Baltimore on EM immunogold studies of several proteins related to protein trafficking in neurons, including distributions in normal rats and knock-in mice. 5) Working with Dr. Jeff Erickson of LSU Health Sciences Center on some EM immunogold studies of synapse plasticity. 6) Working with Dr. Youming Lu of LSU School of Medicine on ultrastructural changes in neurons related to signaling in synaptic transmission.